A Model of Glial Scarring Analogous to the Environment of a Traumatically Injured Spinal Cord Using Kainate

• Published in: Annals of rehabilitation medicine Vol. 40; no. 5; pp. 757 - 768
• Main Authors: Yoo, Jong Yoon, Hwang, Chang Ho, Hong, Hea Nam
• Format: Journal Article
• Language: English
• Published: Korea (South) Korean Academy of Rehabilitation Medicine 01.10.2016; 대한재활의학회
• Subjects: Original; 재활의학; Spinal cord injuries; Kainic acid; In vitro techniques; Neuroglia
• ISSN: 2234-0645; 2234-0653
• DOI: 10.5535/arm.2016.40.5.757

To develop an model analogous to the environment of traumatic spinal cord injury (SCI), the authors evaluated change of astrogliosis following treatments with kainate and/or scratch, and degree of neurite outgrowth after treatment with a kainate inhibitor. Astrocytes were obtained from the rat spinal cord. Then, 99% of the cells were confirmed to be GFAP-positive astrocytes. For chemical injury, the cells were treated with kainate at different concentrations (10, 50 or 100 µM). For mechanical injury, two kinds of uniform scratches were made using a plastic pipette tip by removing strips of cells. For combined injury (S/K), scratch and kainate were provided. Cord neurons from rat embryos were plated onto culture plates immediately after the three kinds of injuries and some cultures were treated with a kainate inhibitor. Astro-gliosis (glial fibrillary acidic protein [GFAP], vimentin, chondroitin sulfate proteoglycan [CSPG], rho-associated protein kinase [ROCK], and ephrin type-A receptor 4 [EphA4]) was most prominent after treatment with 50 µM kainate and extensive scratch injury in terms of single arm (p<0.001) and in the S/K-induced injury model in view of single or combination (p<0.001). Neurite outgrowth in the seeded spinal cord (β-III tubulin) was the least in the S/K-induced injury model (p<0.001) and this inhibition was reversed by the kainate inhibitor (p<0.001). The current model combining scratch and kainate induced glial scarring and inhibitory molecules and restricted neurite outgrowth very strongly than either the mechanically or chemically-induced injury model; hence, it may be a useful tool for research on SCI.